Actinobacillus actinomycetemcomitans is implicated in the pathogenesis of localized aggressive periodontitis, and has the capacity to express a cytolethal distending toxin (Cdt). Gingival fibroblasts (GF) are resident cells of the periodontium, which can express several osteolytic cytokines. The aims of this study were a) to investigate the role of Cdt in A. actinomycetemcomitans-induced expression of osteolytic cytokines and their cognate receptors in GF and b) to determine if the previously demonstrated induction of receptor activator of NFκB ligand (RANKL) by A. actinomycetemcomitans is mediated by these pro-inflammatory cytokines or by prostaglandin E (PGE). A. actinomycetemcomitans clearly induced interleukin (IL)-6, IL-1β, and to a minimal extent, tumor necrosis factor (TNF)-α mRNA expression. At the protein level, IL-6 but not IL-1β or TNF-α expression was stimulated. The mRNA expression of the different receptor subtypes recognizing IL-6, IL-1β and TNF-α was not affected. A cdt-knockout strain of A. actinomycetemcomitans had similar effects on cytokine and cytokine receptor mRNA expression, compared to its parental wild-type strain. Purified Cdt stimulated IL-6, but not IL-1β or TNF-α protein biosynthesis. Antibodies neutralizing IL-6, IL-1 or TNF-α, and the PGE synthesis inhibitor indomethacin, did not affect A. actinomycetemcomitans-induced RANKL expression. In conclusion, a) A. actinomycetemcomitans induces IL-6 production in GF by a mechanism largely independent of its Cdt and b) A. actinomycetemcomitans-induced RANKL expression in GF occurs independently of IL-1, IL-6, TNF-α, or PGE. © 2005 Elsevier Ltd. All rights reserved.